The study of biologically active substances of chaenomelesand the products of its processing

The importance of developing food products of improved biological value to ensure the healthy nourishment of the population was analyzed. The prospects of using plant raw materials as a source of biologically active components were shown. The chemical composition of chaenomeles fruit and of the products of its processing was determined. The biological value of the components of the fruit was explored, and it was found that maximum amount of L­ascorbic acid is contained in the pulp of the fruit and that maximum amount of phenolic substances is contained in the peels of the fruit. With the help of chromatographic studies, it was established that the products of chaenomeles processing have significant content of organic acids, among which malic, quinic, citric and succinic acids were identified, malic acid is dominant amomg them. The sugars, found in the fruit of chaenomeles, are represented by fructose, glucose and saccharose, among them fructose and glucose prevail. The raw material contains procyanidins, hydroxycinnamic acids, flavones and flavan­3­ols, among them procyanidin trimmer, chlorogenic acid, rutin and epicatechine dominate, which have high antioxidant properties. In the products of chaenomeles processing, 48 types of aromatic compounds were identified, among which prevail alcohols, acids, ethers and unsaturated carbohydrates that give products of chaenomeles processing unique aroma and predetermine their antibacterial properties. The products of chaenomeles processing (juice, puree) are a valuable source of organic acids; they were used as a natural regulator of acidity and as an antioxidant in manufacturing products from flour yeast dough. Puree fromchaenomeles contains a significant amount of pectic substances and was used in the production of fruit sauces as a structuring agent. An increase in the organoleptic and physical and chemical indicators in fruit sauces and flour products with the use of products of chaenomeles processing was established.


Introduction
The problem of present time is substantial deterioration of health and a decrease in life expectancy of the population, which is caused by unfavourable environmental situation, low quality food, artificial food additives and consumption of a great amount of refined products. Result of the influence of these factors is disruptionof normal physiological state of a human body, increase in the number of oncologicaland various professional diseases. The most acute problem is the health of children, because every second child is born with certain types of allergic diseases, congenitalabnormalities and other defects of normal development [1].
Over the past decades, the role of biologically active components of foods of different chemical nature has been studied in order to prevent the most dangerous and socially significant chronic non-infectious diseases.
Arelevant task of today is to create food products with a directed biological effect due to the use of natural ingredients of the raw materials that possess antioxidant and adaptogenic properties.
In this regard, it is socially important to develop the technology of food products withimproved consumer prop-erties, which imply a decrease in energetic versus nutritious value and enhancing organoleptic parameters.

Literature review and problem statement
The leading scientists were involved in the development of scientific fundamentals of technology of processing the alternative raw material with its use in manufacturing food products [2,3].
Fruit and vegetable and plant raw materials and products of their processing (fruit and vegetable powders, pastes, purees, concentrates and juices) are a source of natural complex of biologically active substances, which positively affect a human organism. They provide vitamins, mineral substances, phenolic compounds and pectic substances that have a wide range of biological impact (hypotensive and vessel-strengthening, radioprotective, detoxicational, etc.). They are used for imrovingstructural, mechanical and organoleptic properties and enriching food products with nutrients.
A valuable source of biologically active substances is chaenomeles, which contains vitamins, organic acids and pectic substances and has a pleasant aroma [4]. Fruits of chaenomeles are widely used in the Eastern medicine. The antibacterial activity of the fruit in relation to e.coli is confirmed by the scholars from Taiwan University [5]. The glycosides, which are contained in the fruit of chaenomeles, are actively used in the treatment of juvenile and collagen-induced arthritis [6]. The anti-inflammatory and analgetic effects of chaenomeles fruit [7], the antiviral (virus groups A and B) and anticancer effects of extracts of the chaenomeles fruitwere proved [8]. The fruit of chaenomeles inhibit tissue thromboplastin [9], prevent thrombogenesis and can be used by the patients suffering from diabetes II and in medical and prophylactic diets [10].
Analysis of the literature sourcesrevealed that chaenomeles was mainly studied for the selection of new varieties [11]. The prospect of using chaenomeles fruit for medical and prophylactic purposes was explored [12,13]. There are no studies onusing it as a biologically active additive for food products. The studies of the quality indicatorsof chaenomeles and of the products of its processing, and using them infood production technologies are promising.

The aim and the tasks of the study
The aim of the work is studying biologically active substances of chaenomeles fruit and the products of its processing and the substantiation of expediency of their use in the production of food products.
To achieve the set goal, the main tasks that had to be solved were defined: -to explore chemical composition of chaenomeles fruit and the products of its processing; -to carry out chromatography studies to identify the composition of phenolic substances, organic acids, sugars and volatile compounds of chaenomeles and the products of its processing; -to explore the influence of products of chaenomeles processing on the quality of fruit sauces and flour products with their use.

Materials and methods of research into main physical
and chemical indicators of chaenomeles and the products of its processing

1. Standardized methods of research into physical and chemical indicators of chaenomeles and the products of its processing
To carry out the research, the mix ofchaenomeles, gathered in Ukraine, were used.
The object of research was chaenomeles, the juice and puree obtained from it, as well as fruit sauces and flour products produced with the use of products of chaenomeles processing.
To identify the main physical and chemical indicators (dry substances, acidity, vitamin C and phenolic substances), the standard methods were used [14,15].

Results of the research into the content of biologically active substances in chaenomeles and the products of its processing
For the analysis of organoleptic indicators, chaenomeles at the stage of consumption ripeness, gathered in mid-September on the territory of Ukraine, was selected. The fruits differed in color from light-yellow to yellow andhad characteristic sour, slightly tart taste and pleasant saturatedaroma that matches their botanical characteristics [11].
The studies to determine the components of chaenomeles fruit and their biological value are shown in Fig. 1. The analysis of previously conducted research [16] and the data of Fig. 1 indicate that 60 % of the fruit is the pulp, which contains in its composition: organic acids (6,33 %), pectic substances (1,82 %), L-ascorbic acid (248 mg/100 g) and phenolic substances (920 mg/100 g), but not less valuable are the peel of the fruit (17 % of the fruit mass) and a seeds chamber (9 % ofthe fruit mass), which contain a significant amount of essential oils and phenolic substances (1400 mg/100 g).
A significant part of biologically active substances (BAS) of chaenomeles are in a combined state, only a part of them is in cell sap and, in the processing, it passesto a soluble part. The main carbohydrates that make up the primary cell membrane are cellulose, hemicellulose and pectin. Cellulose microfibers are connected through hemicellulose bridges, forming a cellulose-hemicellulose network, which is surrounded by the matrix of pectin. Phenolic compounds are mainly localized in the peel and in thecell wall of the pulp of raw material and are an obstacle totransfer of useful ingredients of the fruit to soluble part. That is why it is expedient to break the integrity of these natural biopolymers by mechanical or chemical action.
The most efficient way of preliminary processing of fruit and berries is the use of enzyme preparations. Their application allows accelerating technological processes, increasing the yield of juice from the raw materials and increasing its nutritional value by enriching with phenolic compounds This method of processing provides an opportunity not only to increase the yield of the juice and extract BAS from the peel and pulp of the fruit, but it is also the most rational method of direct extraction for enterprises in public catering [7].
The conducted experimental studies revealed that the highest yield of puree and the lowestvolume of waste is achieved when blanchingchaenomeles fruit in water. This method of preliminary preprocessing is the most affordable and rational under conditions of restaurant business [18].
Physical and chemical characteristics of quality were definedin chaenomeles fruits and in the products of their processing (juice and purée), the results of which are shown in Table 1. Table 1 Physical and chemical indicators of quality of chaenomeles and theproducts of its processing (n=3, p≤0,05) The results, shown in Table 1, prove that the products of chaenomeles processing contain a significant fraction of pectic substances, phenolic compounds and L-ascorbic acid in their composition.
Pectic substances increase resistance of organism to allergies, contribute to restoration of the mucose membrane of respiratory and digestive tracts after diseases, affect the overall metabolism and are immunity strengthening means. Pectic substances are found almost in all plants. The content of pectic substances in chaenomeles fruit is 40 % higher than that of apples. Prophylactic rate of pectic consumption is 2 g per day [19].
Chaenomeles contain 6 times more of L-ascorbic acid, known for its antioxidant properties, than lemons [4]. The daily need for ascorbic acid for adults is 70-80 mg per day [20]. In the course of processing chaenomeles fruits, 42 % of L-ascorbic acid are lost while making puree and up to 60 % of it is lost while making juice, although its content remains relatively high.
To determine biological value of the products of chaenomeles processing, the study was conducted of the composition of organic acids and sugars ( Table 2).

Тable2
Composition of organic acids and sugars in the products of chaenomeles processing (n=3, p≤0,05) It can be seen from the results of the study ( Table 3) that sugars, which are represented exclusively by hexoses -glucose and fructose, are the main fraction of soluble dry substances in chaenomeles. Fructose is the sugar that is easily assimilated by organism, so the products of chaenomeles processing can be used in the technology of food products for dietary purposes, including the diet of people who suffer form diabetes.
In addition to sugars, taste properties of raw material are determined by the presence of organic acids. It was found that in the products of chaenomeles processing they are mainly represented by malic and quinic acids, as well as citric and succinic acids, which increases antioxidant properties of the fruit.
Availability of considerable amount of malic acid will allow using the products of chaenomeles processing as a natural regulator of acidity in foods.
Quinic acid is an important intermediate product of biosynthesis of aromatic compounds (flavonoids, phenolcarbonic acid and others) in higher plants and some microorganisms and has positive effect on a human organism.
Succinic acid can act as a reducing and radical-accepting agent, responsible for antioxidant protection. The main effect of succinic acid on a cell as an antioxidantis in reducing the intensity of the flow of lipid peroxidation, in increasing the content of restored glutathione, in reducing thiol-disulfide status of the cell and enhancing activity of antioxidant enzymes [21].
Phenolic compounds play a role of inhibitors in the process of oxidation of BAS. The specificity of the influence of certain phenolic compounds on reactivity of the components of crushed fruit was used to decrease the degree of oxidative transformations in them.
The research into the fractional composition of phenol substances contained in the products of chaenomeles processing (Table 3) show a significant content of procyanidins that have antioxidant activeness, which is 20 times higher than ascorbic acid and 50 times higher than vitamin E. Procyanidins contribute to the increase in macromolecular lipoproteins in blood plasma, which leads to reducing the risk of cardio-vascular diseases [22], and, as a reducing substance, is involved in preventing cancer diseases of gastro-intestinal tract and internal organs [23]. In addition, procyanidins promote weight loss and prevent development of diabetes of type 2 [24]. Catechins and chlorogenic acid also possess antioxidant properties; besides, catechins have antimicrobial properties and are used for strengthening immunity.
Rutin is an important substance in prevention and treatment of hypovitaminosis and vitamin P deficiency in the diseases that are accompanied by disruption of vascular permeability and allergic diseases.
Given that the semi-finished products from chaenomeles possess potent antioxidant properties, we can stresshigh biological value of food products made with their use.
The research into fractional composition of volatile substances (Table 4) in the products of chaenomeles processing prove its considerable biological value.

Тable3
Composition of phenolic substances in the products of chaenomeles processing (n=3, p≤0,05) The obtained data prove that the aroma of juice and puree is formedby aromatic alcohols, acids, aldehydes, ketones, ethers, carbohydrates that are present in raw materials. A significant amount of volatile aromatic substances are found in puree (Fig. 2, 3) sincea whole fruit is used in its preparation, including the peels -the main source of aromatic substances.
The research results (Table 4) prove that acids, ethers and carbohydrates prevail among volatile compounds, which are represented in the products of chaenomele processing.
Alcoholsare represented by unsaturated, saturated and aromatic alcohols. The combination of α-terpeniol (lilac), β-terpeniol (hyacinth), β-ionone (violet), α-farnesene (green apple), estragole (dragon), lianole (lily-of-the-valley), eudesmole (rose, eucalyptus), and ethylcaprylate (flower aroma) providechaenomeles and the products of its processing with unique strong fragrance. Special attention should be paid to the presence of squalene (representative of unsaturated carbohydrates) in the amount of 2,11 mg/kg in the juice and 6,43 mg/kg in the puree; it is considered "a vitamin of oxygen" and is a unique component of the skin lipids of a human. In many countries of the world,the preparation of squalene is used in the treatment of atherosclerosis, coronary heart disease, high cholesterol, liver and skin diseases, so its availability inthe products of chaenomeles processing increases its biological value.
Upon analysis of the data from Table 4, it was noted that acids make upalmost 60 % of the total content in puree and 30 % in juice. The group of acids is largely represented by carboxylic acids, among which the most important are caprylic and lauric acids, which have antimicrobial and antiseptic properties, immunity strengthening effect and help fight diabetes and high blood pressure. There can be found linoleic and oleic acids, known as ω-6 and ω-9 unsaturated aliphatic acids that have positive impact on a human organism and are used for prevention of diseases of different etymology [26].
Antimicrobial and antibacterial properties of the products of chaenomeles processing make it possible to predict a decrease in microbiological contamination of foods with the use of chaenomeles.
Given nutritional and biological value of the products of chaenomeles processing, it is rational to use them in technology of sweet sauces and pastry products, with the aim of improving organoleptic, structural and mechanical, physical and chemical indicators of the finished products.
Physical and chemical quality indicators of the finished products are presneted in Tables 5, 6.
The results of the research (Table 5) prove that the use of the products of chaenomeles processingin sweet sauces enhances their biological value by increasing the content of L-ascorbic acid, pectic and phenolic substances compared with the reference sample.  Consequently, adding products of chaenomeles processing to the formulations of flour products from yeast dough ( Table 6) increases porosity ofthe finished products, which contributes to their better digestibility. An increase in humidity and acidity in the experimental samples, compared with the reference samples, allows increasingterms of storage of the finished products and preventing development of potato disease.

Тable 6
Physical and chemical indicators of quality of finished products made of yeast dough usingthe products of chaenomeles processing (n=3, p≤0,05) 6. Discussion of the results of studying the content of biologically active substances in chaenomeles and the products of its processing The conducted studies ofdetermining chemical composition of chaenomeles and the products of its processing prove the feasibility of their use in the production of food.
A valuable source of organic acids is chaenomeles juice, which should be used as a natural regulator of acidity and as an antioxidant. Availability of amino acids, mineral substances and monosaccharides in the juice enriches the flour medium and createsoptimal conditions for the yeast vitality, shortening the process of dough formation. The set of aromatic substances contained in chaenomeles juice addsa pleasant aroma to ready products [27].
In addition, chaenomeles juice can be used as a natural inhibitor of oxidative reactions that occur in plant raw materials under the influence of polyphenoloxidase that is present in Jerusalem artichokein large amount, which complicates its processing. Therefore, the pre-treatment of Jerusalem artichoke in chaenomeles juice is proposed to prevent its darkening before obtaining puree that is used as a semi-finished product in the technology of sauces [18].
One of the directions of using chaenomeles juice is its use for softening the tissue of plant raw materials, which include pumpkin. It was found that the most efficient is the pre-treatmentof such raw material in citric acid and incheanomeles juice, which contains malic, quinic and tartaric acids, with subsequent blanching in water. Organic acids contained in chaenomeles juice positively affect not only the oxidation reactions that take place in pumpkin but also its organoleptic characteristics. Since chaenomeles puree combines considerable content of pectic substances and organic acids, it can be used as a natural structuring agent in the technology of fruit sauces and pastry products. The use ofchaenomeles puree in the technology of flour products from yeast dough, made of flour with weak gluten, contributes to strengthening the dough structure and the structure of gluten proteins, which can be explained by oxidation of proteins under the influence of L-ascorbic acid, organic acids and by formation of complexes with polyphenols and pectins contained in puree in sufficient quantities.
Through considerable acidity, it is not expedient to use chaenomeles puree in its pure form, that is why, in the course of developing the sauces formulations, it was blended with other pectin containing raw materials with the highest sugar-acid index: apples, pumpkin and Jerusalem artichoke. The developed sauces have high structural and mechanical properties, temperature stability and high nutritional value [18].
In addition, the use of semi-finished chaenomeles products in food technology reduces their overall microbiological contamination, prevents development of potato diseases, which is achieved due to the content of certain carbonic acids and is proved by the research.
Along with it, the use of chaenomeles products in food products provides an opportunity to enhance their biological value and antioxidant properties due to the content of phenolic substances and L-ascorbic acid.
2. According to the results of chromatographic studies, the composition of phenolic substances in the products of chaenomeles processing was identified. It was determined that the juice and puree contain procyanidins,

Тable 5
Physical and chemical indicators of sweet sauces made of the products of chaenomeles processing (n=3, p≤0,05) hydroxycinnamic acids, flavones and flavan-3-ols. The dominant representatives are procyanidin trimmer, chlorogenic acid, rutin and epicatechine. Among the organic acids, malic, quinic, citric and succinic acids were identified, malic acid being dominant among them. Sugars are represented mostly by fructose and in a less degreeby glucose and saccharose. Among the volatile compounds that characterize fragrance of the products of chaenomeles processing, 51 compounds were found and 48 compounds were identified.
3. It was established that the use of products of chaenomeles processing affects quality indicatorsof fruit sauces and flour products with their use: in sauces, the content of L-ascorbic acid, pectic and phenolic substances increases; in flour products with yeast dough, porosity, specific volume, resistance to form changing, humidity and acidity increase.
Based on the conducted research, it was substantiated that chaenomeles is a valuable raw material and has prospects for its use in the manufacture of food products.